A Alcaligenes strain ho-1 and its application
A technology of HO-1 and alcaligenes, which is applied in the direction of bacteria, biological water/sewage treatment, sustainable biological treatment, etc., can solve the problems of cumbersome operation, low organic load tolerance threshold, slow reactor start-up, etc., and achieve simplification Denitrification process flow, high-efficiency ammonia nitrogen removal capacity, and the effect of saving treatment space and cost
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Embodiment 1
[0026] Example 1. Identification of bacterial strain HO-1
[0027] The Alcaligenes sp. strain HO-1 of the present invention is isolated from a short-path nitrification reactor for wastewater treatment in pig farms.
[0028] Inoculate strain HO-1 on LB solid slant medium, cultivate at 30°C until obvious bacterial lawn appears, pick a small amount of bacterial lawn with a sterile inoculation loop and mix it in an EP tube filled with 200 μL sterile water, vortex Mix to make a bacterial suspension. Then put the EP tube into the boiling water bath and cook for 3 minutes, and put it on ice immediately after taking it out. Bacteria cells are prone to rupture after transient hot and cold alternation, and then release genomic DNA. The lysed bacterial suspension was boiled and centrifuged for a short time, and the supernatant was taken out, which could be used as a template for subsequent 16S rRNA gene PCR amplification.
[0029] The bacterial universal primer 27F / 1492R (27F: 5'-AGAG...
Embodiment 2
[0036] Embodiment 2. The growth and ammonia nitrogen removal of bacterial strain HO-1 with ammonia nitrogen as the only nitrogen source
[0037] Inoculate the HO-1 strain cryopreserved in glycerol into a test tube containing LB liquid medium, culture it in a shaker at 30°C and 160rpm for 24h to activate it, and then transfer 1% of the inoculum into a test tube containing 100mL LB liquid medium Cultivate in a 250mL Erlenmeyer flask at 30°C and 160rpm for 20h to be a seed culture solution.
[0038] The seed culture solution was centrifuged at 6000 rpm for 5 minutes to collect the cells, washed with sterile normal saline for 3 times, and then resuspended with an equal volume of sterile normal saline to prepare a seed suspension.
[0039] The above seed suspension was inoculated into a 500mL Erlenmeyer flask containing 200mL of HNM medium at an inoculation amount of 1.5%, and cultured at 30°C and 160rpm for 48h. Take out 3mL culture medium regularly, 1mL is used for measuring the...
Embodiment 3
[0046] Example 3. Detection of Nitrogen Products in the Denitrification Process of Bacterial Strain HO-1
[0047] Activate the bacterial strain HO-1 according to the method in Example 2, take 4 mL of seed culture solution, centrifuge at 6000 rpm for 5 minutes to collect the bacteria, wash the bacteria three times with sterile normal saline, and then resuspend with 8 mL of normal saline to prepare 2-fold diluted seeds Suspension.
[0048] Use a 2mL syringe to inoculate 1.2mL of the above seed suspension into a 500mL airtight bottle (not ventilated) containing 40mL nitrogen detection medium. Shaker culture. After 4 days, take the gas in the bottle and use the method of stable isotope ratio mass spectrometry to detect whether stable isotope-labeled nitrogen has been produced.
[0049] Nitrogen detection medium formula (g / L): 15 N stable isotope labeled NH 4 Cl 0.19, sodium hexahydrate 22.51, KH 2 PO 4 0.5, Na 2 HPO 4 12H 2 O 1.25, MgSO 4 ·7H 2 O 0.2, trace element sol...
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